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`West-Ward Exhibit 1054
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`West-Ward Exhibit 1054
`Morris 1989 - Page 002
`
`

`

`,',;fed, Sci. Res"
`
`17,609-610
`
`609
`
`anew
`
`the survival
`
`(RPM), a novel
`developed as an
`of organ grafts
`strains of 1'0-
`
`We report for the first time that
`macrolide fermentation
`agent. prolongs
`between highly
`dents without apparent
`As
`result of largely
`respectability and
`decades, transplantation has
`is now the
`treatment for several serious diseases,
`recent advances. however, transplantation is an
`therapy that is far from
`its full potentiaL
`transplantation of cadaveric
`is still an expensive,
`labour-intensive treatment for
`relatively small number of
`who still must endure an unacceptably high incidence
`of morbidity and mortality related to rejection. infection,
`and drug
`that prevents complete rehabi-
`litation for these patients. The best that transplantation can
`offer is palliation - not a cure. All of the
`that
`continue to haunt the field
`root cause: our
`immune response to the
`and inexpensively.
`of immunosuppressive
`both the central
`drug development to continued progress in transplantation
`and the rapid accumulation of our knowledge of the immune
`system, it is noteworthy that clinical transplantation must
`only four drugs
`of which were introduced a
`generation ago) to control
`of identifiying new lead
`into clinically
`our laboratory has
`compounds. FK506
`sive compound that we [l J and others
`investigate. Although its immunosuppressive actions are
`similar to cyclosporine
`FK is a macrolide and is.
`therefore. structurally distinct from CsA. It is still too
`to
`predict what role this new, potent and very effective agent
`will play in clinical transplantation since it is still in the
`formative stages of development.
`at
`mid-
`RPM
`was discovered in
`Wyeth-Ayerst Research Lahoratories. found to have antifun-
`gal activity and, like
`is a macrolide fermentation
`As part of routine animal
`studies.
`of RPM were found to alter lymphoid tissue
`subsequent studies in rats showed that RPM was also an
`autoimmune
`effective treatment of
`disease
`. This immunosuppressive
`of RPM discour-
`aged its
`as an antifungal agent, and although
`there was some interest in
`its anti tumour activity.
`the
`molecule remained a potentia! solution In search of an
`appropriate clinical problem,
`Several lines
`evidence made a
`" .. v.r'A~ of RPM's utility as an
`is stucturally
`T-cell mediated
`rejection:
`similar to FK and FK is a proven inhibitor of T-cell activation
`and graft
`RPM suppresses T-cell mediated
`RPM appears to have a wide
`since the acute
`dose of RPM in rats is
`margin of
`many times more than the dose of RPM that suppresses
`autoimmune diseases in this species
`5].
`To learn whether the treatment t,f
`
`recipients with
`
`the survival of transplanted
`RPM was capable of
`tissue, two murine models of organ transplantation were
`used. In the first modeL we determined the survival times of
`donor BALB/c
`mouse neonatal hearts transplanted
`into the car pinnae of
`C3H/Km (H-2k) mice. The
`was a modification of the method we have used to
`cyclosporine
`The
`control group consisted of untreated and saline
`treated animals. Recipients in the RPM treatment group were
`treated
`from post
`day I through 13 by gavage
`with powdered RPM (provided by
`Research, Princeton,
`in 2% carboxymethyl-
`cellulose in water. Each heart graft was examined
`every other day from
`6 until day 14 and then
`Wednesday and
`until the day of rejection
`survival) which was defined as the first
`contractile
`was seen.
`rat hearts
`In the second model. Brown Norway
`were transplanted
`vascular anastomoses into the
`abdamens of Lewis
`rat recipients as
`described rR],
`in the control group were treated
`with normal saline intraperitoneally (IF)
`from post
`operative day 1 until the grafts rejected.
`in the
`treatment group were treated daily by gavage with RPM
`formulated a~ described above from post
`day 1 .. 1-, Each heart
`was palpated daily until the
`rejection which was defined as the day on which graft
`contractile
`had ccased as confirmed
`at
`
`on which no
`
`purposes, mouse and rat
`certain groups were treated
`with cv,CIC,SDon
`oiive oil
`a treatment schedule identical to that used for
`RPM. In both murine models the donors and
`are
`incompatible across the major histocompatibility complex.
`The
`U-test with a correction for small sample
`sizes was used to determine the statistical significance of the
`differences in
`survival times between groups.
`Table 1 shows the results of the
`of the initial
`lise of RPM as an immunosuppressant for
`rejection. An
`oral
`of mg
`of RPM administered for only the first
`13
`days enabled BALB/c hearts transplanted
`to survive a median of 15
`with the graft survival times in the saline/no
`treatment group, the prolongation of
`survival in mice
`(p < O.OOOI), Even
`treated
`RPM was highly
`CsA was significantly (p <
`less effective than
`the dose of RPM that was half the CsA prolonged
`survival significantly compared with the control group, CsA
`dose,
`
`to the anti rejection
`were also
`in the RPM-treated
`since the heart
`survived significantly
`0.001) longer than
`in
`those animals treated with saline. Unlike OUf results in the
`mouse. we did not find that RPM was a more potent
`immunosuppressant than CsA for rat allograft
`Micronization of the RPM. formulation in an
`alternative routes may
`and so
`increase
`its
`
`increase
`potency,
`Since we are at such an
`
`stage in our evaluation of the
`
`West-Ward Exhibit 1054
`Morris 1989 - Page 003
`
`

`

`610
`
`Le!lers
`
`Table j: Prolong{/[ion of heart graft survival by orally administered (RPl'vf) and cyclosporine (CsA).
`
`Group survival times
`
`Species Treatment
`
`Dose
`(mg kg ')
`
`fndividual survival times
`( days)
`
`Mean
`
`Median
`
`p values
`
`Mouse" Saline
`
`Mouse
`
`CsA
`
`Mouse
`
`RPM
`
`Rill b
`
`Saline
`
`Rat
`
`Rat
`
`CsA
`
`RPM
`
`12
`
`6
`
`2
`
`3
`
`1O( x74), 12( x29), 14( x2)
`
`10,6 ± 0.2
`
`8,10, 12( x5), 14
`
`to, 12, 12, 14, 14, 16( x4), 19
`
`6,7(x6)
`
`9.24,24,30
`
`15,22,22,29,31
`
`12 ± 1.5
`
`14 ± 1.9
`
`7
`
`0.35
`
`22 ± 14
`
`24 ± 7.9
`
`10
`
`12
`
`15
`
`7
`
`24
`
`22
`
`"BALBic /() C3fJ; "Brown Norway to Lewis; cMann, Whitney V-lest.
`
`=0,026
`
`=0.013
`
`=0.003
`
`=1.0
`
`<0.0001
`
`=0,001
`
`Randall Ellis Morris
`Bruno Michael Meiser
`
`Laboratory for Transplantation
`Immunology, Department of
`Cardiovascular Surgery, Falk
`Cardiovascular Research
`Stanford University
`School of Medicine Stanford, CA
`94305-5247, USA.
`
`We commend Suren Seghal's interest in RPM's potential use
`as an immunoregulator for graft rejection and acknowledge
`him for bringing this molecule to our attention, We also
`gratefully acknowledge the expert technical assistance of
`Randi Shorthouse, Paula O'Conner and Jun Wu, This work
`was supported by the Peyton Hawes and Hedco foundations
`and by a NATO Fellowship awarded to RM,M,
`
`Note added in proof: We have learned from Wyeth-Ayerst
`that when R. Y. Calue's group in Cambridge,
`was
`provided with RPM, they too found that the compound
`prolonged the survival at allografts in animals.
`
`immunosuppressive activity of RPM, few conclusions can be
`made with certainty, The data do show that RPM prolongs
`in mouse
`the survival of highly histoincompatible heart
`and rat transplant recipients, As the acute
`for orally
`administered RPM are so high for the mouse and rat
`it
`was not surprising that the doses of RPM that we used to
`prolong graft survival caused no noticeable signs of toxicity,
`RPM is now the second macrolide to show sufficient
`immunosuppressive activity to prolong the survival of trans(cid:173)
`planted tissue, The true mechanisms of immunosuppressive
`action of RPM are not known, but it is tempting to speculate
`that RPM and FK control the rejection response in similar
`ways if only because they share certain similarities in their
`molecular structure, Ultimately, the clarification of the
`contrasting mechanisms of immunosuppressive action of
`these macrolides and CsA may allow us to understand why
`these fungal fermentation products are so specifically suited
`to the control of the mammalian immune response,
`In a more practical vein, further investigation of RPM is
`needed to define its potency, efficacy, selectivity and safety
`so that its potentia! for clinical use can be assessed and
`compared to the drug profiles of other known immunosup(cid:173)
`pressive agents. Comprehensive studies of this nature are
`ongoing in our laboratory; we now have evidence that
`transient treatment with RPM at doses higher than the doses
`used here or given by different routes induces indefinite
`unresponsiveness in recipients.
`The route from the discovery of a new lead compound to
`treacherous and tortuous.
`the final clinical use is often
`Although the trail leading to the use of RPM as an antifungal
`agent has grown cold, the fresh scent of this compound's
`anti rejection activity deserves attention. The apparent safety
`of the drug at doses that are immunosuppressive is reason for
`cautious optimism that RPM may contribute in some way to
`the solution of the serious problems that continue to
`clinical transplantation.
`
`I. Morris, R,E" lIoy!, G" Murphy, M,P, et al. 1989, Transplam. Pmc., 21,
`1042,]()44
`Thomson, A.W. 19!N. Immunology
`2.
`3. Vezina, C. Kudclski. A. and Sehgal,
`721-726
`4 Manel, R.R., Klicius, J. and Gale!, S. 1977. Can. 1. Physiol, Pharmacol.,
`55, 48,51
`Sehgal, S,N" Wyeth-Ayers! Research, Princeton, NJ, USA, (personal
`communicatlon).
`Batmy, G., Morris, R.E" Babny, L el at. 1988,1, Pharm, Exp, Ther"
`164, 24-58
`Babny, G" Morris, R.E., Babny, L el ai, 1989.1. Pharm, Exp. Ther.,
`248, 89],899
`8. Morns, R.E. and Meiser, B.M. 1988. Transplant. fmc.,
`Baker, H .. Sldcrowicz. A., Sehgal, S.N, et al. 1978,1.
`9,
`539-545
`
`HI, 6-9
`1975. I Antibiotics, 28,
`
`659-660
`31,
`
`5,
`
`7.
`
`Letter received: 27(h May, 1989; amended 13(h June, 1989.
`
`West-Ward Exhibit 1054
`Morris 1989 - Page 004
`
`